Wholemeal flours from 62 spelt and 13 wheat cultivars were studied. The quantitative protein compositions of the Osborne fractions determined by reversed-phase high-performance liquid chromatography, showed that the chromatograms of the reduced gliadin fractions were most suitable for the distinction of spelt from wheat and for the classification of spelt. The patterns of the reduced spelt gliadins showed one to three markers that were not present in wheat. Based on these markers, spelt cultivars were classified into three groups ranging from typical spelt to similar to common wheat. Marker 1 was identified as 1,2-gliadin and markers 2, 3a and 3b were identified as -gliadins by means of N-terminal sequence analysis and determination of the relative molecular mass by mass spectrometry. As glutenin-bound -gliadins were present in wheat and absent in spelt, this protein type may be used to detect and quantitate small amounts of wheat in spelt products.